WO2006127967A3 - Multifocal scanning microscopy systems and methods - Google Patents

Multifocal scanning microscopy systems and methods Download PDF

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Publication number
WO2006127967A3
WO2006127967A3 PCT/US2006/020372 US2006020372W WO2006127967A3 WO 2006127967 A3 WO2006127967 A3 WO 2006127967A3 US 2006020372 W US2006020372 W US 2006020372W WO 2006127967 A3 WO2006127967 A3 WO 2006127967A3
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WO
WIPO (PCT)
Prior art keywords
mapmt
raster
specimen
methods
mmm
Prior art date
Application number
PCT/US2006/020372
Other languages
French (fr)
Other versions
WO2006127967A2 (en
Inventor
Karsten Bahlman
Ki-Hean Kim
Timothy Ragan
Peter T C So
Original Assignee
Massachusetts Inst Technology
Karsten Bahlman
Ki-Hean Kim
Timothy Ragan
Peter T C So
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Massachusetts Inst Technology, Karsten Bahlman, Ki-Hean Kim, Timothy Ragan, Peter T C So filed Critical Massachusetts Inst Technology
Priority to EP06784478A priority Critical patent/EP1889111A2/en
Publication of WO2006127967A2 publication Critical patent/WO2006127967A2/en
Publication of WO2006127967A3 publication Critical patent/WO2006127967A3/en

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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/645Specially adapted constructive features of fluorimeters
    • G01N21/6452Individual samples arranged in a regular 2D-array, e.g. multiwell plates
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/645Specially adapted constructive features of fluorimeters
    • G01N21/6456Spatial resolved fluorescence measurements; Imaging
    • G01N21/6458Fluorescence microscopy
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64Fluorescence; Phosphorescence
    • G01N21/6486Measuring fluorescence of biological material, e.g. DNA, RNA, cells
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/0004Microscopes specially adapted for specific applications
    • G02B21/002Scanning microscopes
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/0004Microscopes specially adapted for specific applications
    • G02B21/002Scanning microscopes
    • G02B21/0024Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/0004Microscopes specially adapted for specific applications
    • G02B21/002Scanning microscopes
    • G02B21/0024Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052Optical details of the image generation
    • G02B21/0076Optical details of the image generation arrangements using fluorescence or luminescence
    • GPHYSICS
    • G02OPTICS
    • G02BOPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00Microscopes
    • G02B21/16Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes

Abstract

In the systems and methods of the present invention a multifocal multiphoton imaging system has a signal to noise ratio (SNR) that is reduced by over an order of magnitude at imaging depth equal to twice the mean free path scattering length of the specimen. An MMM (multi-photon multi-focal microscopy) system based on an area detector (120A) such as a multianode photomultiplier tube (MAPMT) that is optimized for high- speed tissue imaging. The specimen (105A) is raster- scanned with an array (151A,152A) of excitation light beams. The emission photons from the array of excitation foci are collected simultaneously by a MAPMT and the signals from each anode are detected using high sensitivity, low noise single photon counting circuits. An image is formed by the temporal encoding of the integrated signal with a raster scanning pattern. A deconvolution procedure taking account of the spatial distribution and the raster temporal encoding of collected photons can be used to improve decay coefficient . We demonstrate MAPMT-based MMM can provide significantly better contrast than CCD- based existing systems.
PCT/US2006/020372 2005-05-25 2006-05-25 Multifocal scanning microscopy systems and methods WO2006127967A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP06784478A EP1889111A2 (en) 2005-05-25 2006-05-25 Multifocal imaging systems and methods

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US68460805P 2005-05-25 2005-05-25
US60/684,608 2005-05-25

Publications (2)

Publication Number Publication Date
WO2006127967A2 WO2006127967A2 (en) 2006-11-30
WO2006127967A3 true WO2006127967A3 (en) 2007-03-08

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US (2) US20070057211A1 (en)
EP (2) EP2703871A3 (en)
WO (1) WO2006127967A2 (en)

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